This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Cells accumulate organic osmolytes when cultured in high osmolality media. Recent evidence indicates that wild type E. coli (eg uropathgenic strain CFT073) possess osmoregulatory transporters and accumulate osmolytes that have not become evident from studies of E. coli K-12. These observations are being pursued using NMR and Mass Spectrometries in an effort to identify previously unknown osmolytes that accumulate to high levels in osmotically stressed bacteria that lack known osmoregulatory systems.